MicroRNA-494-3p targets CXCR4 to suppress the proliferation, invasion, and migration of prostate cancer.

BACKGROUND: Although SDF-1/CXCR4 pathway is a potential mechanism of tumor proliferation and progression, the mechanism of controlling CXCR4 expression is not fully understood. This study was to confirm that miR-494-3p might be a potentially post-transcriptional regulator of CXCR4 and over-expression of miR-494 might suppress prostate cancer progression and metastasis. MATERIALS AND METHODS: We firstly postulated the post-transcriptional regulation of CXCR4 by miR-494-3p through bioinformatics analysis, and then it was demonstrated that miR-494-3p could regulate the CXCR4 mRNA post-transcriptionally by binding to the predicted site by dual reporter gene assays. The biological effect of miR-494-3p on prostate cancer cells proliferation, apoptosis, migration, and invasion was measured by MTT, TUNEL, flow cytometry, migration, and invasion assays. RESULTS: It was shown that the mRNA and protein expression levels of CXCR4 were significantly up-regulated in PC-3 and DU145, whereas barely detected in LNCaP and RWPE-1. However, the CXCR4 protein levels were inversely related to the mature miR-494-3p expression levels in RWPE-1 and prostate cancer cells. The constitutive over-expression of miR-494-3p could down-regulate the protein level of CXCR4 in PC-3 and DU145. MiR-494-3p also could bind to the seed sequences in the 3'-UTR of the CXCR4 gene. Artificial over-expression of miR-494-3p could inhibit the growth, promote the apoptosis, and inhibit the migration and invasion of PC-3 and DU145 cells in vivo. CONCLUSIONS: Our results suggested that miR-494-3p might play crucial role in prostate cancer by post-transcriptional regulation to CXCR4 mRNA. MiR-494-3p/CXCR4 pathway may be a potential therapeutic target to prevent prostate cancer progression and metastasis.

Assessment of the effects of induced anisometropia on binocularity with glasses-free 3D technique.

AIM: To assess the effect of experimentally induced anisometropia on binocularity in normal adults with glasses-free three-dimensional (3D) technique. METHODS: Totally 54 healthy medical students with normal binocularity in the cross-sectional study were enrolled. Anisometropia was induced by placing trail lenses over the right eye, in 0.5 D steps including lenses of -0.5, -1, -1.5, -2, -2.5 D (hyperopic anisometropia) and lenses of +0.5, +1, +1.5, +2, +2.5 D (myopic anisometropia). The glasses-free 3D technique was used to evaluated not only fine stereopsis, but also coarse stereopsis, dynamic stereopsis, foveal suppression, and peripheral suppression in these subjects. One-way analysis of variance was used to compare quantitative data such as fine stereopsis, coarse stereopsis. Pearson's Chi-square test was performed to compare categorical data such as dynamic stereopsis, foveal suppression and peripheral suppression. RESULTS: The subjects showed a statistically significant decline in fine stereopsis, coarse stereopsis, and dynamic stereopsis with increasing levels of anisometropia (P<0.001). Binocularity was affected when induced anisometropia was more than 1 D (P<0.05). Foveal suppression and peripheral suppression were evident and increased in proportion to anisometropia (P<0.001). CONCLUSION: The relatively low degrees of anisometropia may have a potentially significant effect on high-grade binocular interaction. The mechanisms underlying the defect of binocularity seem to involve not only foveal suppression, but also peripheral suppression.

[Clinicopathologic observation of orbital natural killer-T cell lymphoma].

OBJECTIVE: To investigate clinical manifestation, diagnosis and treatment of orbital natural killer (NK)-T cell lymphoma. METHODS: It was a retrospective case series. Seven orbital NK-T cell lymphoma patients confirmed by surgical biopsies were collected during the past 22 years. We reviewed the records, surgical and treatment procedures. Surgical specimens were studied with HE staining, immunohistochemical staining and molecular biological analysis. RESULTS: These patients had proptosis, eye motive inhibition or fixation and visual acuity was decreased or even without light perception. Skin of inner canthus and eyelids appeared red and swollen, with ulceration and cavity formation. CT scan revealed that the tumor showed uneven density and an unclear border. Tremendous lymphocyte infiltration and tissue necrosis in the tumor were observed in the biopsy tissue. LCA, CD45RO and CD57 immunohistochemical staining revealed positive results. Clonal T-cell-receptor gene rearrangements of two patients showed negative results and the Epstein-Barr virus was detected. CONCLUSIONS: Orbital NK-T cell lymphoma is a rare disease. The characteristics of this disease include a highly aggressive clinical course, severe destruction and a poor prognosis. The final diagnosis depends on HE staining, immunohistochemical staining and molecular biological examination.

[Constructing recombinant plasmid with prostate-specific membrane antigen promoter and reverse enhancer and comparing the transcription activity of enhancers in different directions].

OBJECTIVE: To assess the specificity of prostate-specific membrane antigen(PSMA) promoter and enhancer in controlling gene expression and to compare the activity of enhancers in different directions for choosing the most suitable prostate-specific PSMA controlling elements. METHODS: PSMA enhancer gene was amplified with PCR, then the enhancer gene was subcloned into the expressing vector pEGFP-PSMA(Pro) reversely to construct the recombinant plasmid pEGFP-PSMA(E(r)-p), which was transfected into different cell lines such as LNCaP, PC-3,MCF-7,A549. Green fluorescent protein (GFP) expression was observed and compared to other recombinants constructed previously. RESULTS: The recombinant plasmid with reverse enhancer was successfully constructed, the PSMA promoter and enhancer showed modulating activity in PSMA-expressed cell line uniquely. PSMA enhancer could increase 30-fold transcriptional activity over the basal level achieved by PSMA promoter alone, and no impact of the direction on the activity of enhancer was noted. CONCLUSION: PSMA promoter/ enhancer is specific to PSMA-expressed cells. The transcriptional activity of reverse enhancer is similar to that of enhancer. PSMA promoter/enhancer has the potential for use in targeted gene therapy of prostate adenocarcinoma.

[The construction of recombinant plasmid with prostate-specific membrane antigen promoter controlling reporter gene expression].

OBJECTIVE: To study the specificity of prostate-specific membrane antigen (PSMA) promoter in controlling gene expression. METHODS: PSMA promoter gene was amplified with PCR, and then the promoter gene was cloned into the vector pEGFP-1 to construct a recombinant plasmid, which was transfected into different cell lines such as LNcap, PC-3, MCF-7, A549. Green fluorescent protein (GFP) expression was observed. RESULTS: The recombinant plasmid constructed, and PSMA promoter uniquely showed modulating activity in PSMA positive cell line. CONCLUSION: PSMA promoter possesses PSMA positive cell specificity, as well as prostatic tissue specificity. PSMA promoter may have the potential for use in targeted gene therapy of prostate adenocarcinoma.

[Use of indocyaine green staining technique for phacoemulsification in white cataract].

OBJECTIVE: To evaluate the efficiency and safety of indocyaine green (ICG) staining technique using in white cataract phacoemulsification. METHODS: In a prospective clinical study, 105 eyes with white senile cataract were divided into two groups: capsular-staining group using ICG (n = 55) and noncapsular-staining group (n = 50). The time for performing the circular continuous capsulorrhexis (CCC), the number of radial tears of anterior capsule, the number of the rupture of capsular bag and the incidence of IOL in-the-bag fixation during the operation were recorded. We also measured the intraocular pressure and observed the changes of corneal endothelial cells, the anterior chamber flare and the keratic precipitation before surgery, 1 day and 3 months after surgery, respectively. The results of these two groups were compared. The quantitative results were analyzed with two-tailed student's t-test, while the qualitative results were analyzed with chi-square test. RESULTS: In operations performed with ICG staining technique, the CCC could be performed more quickly, no radial tears of the anterior capsule or rupture of capsular bag occurred and all IOL could be fixed in the bag. No statistically significant difference could be detected in the occurrence of these four aforementioned changes between these two groups. None of these patients had an elevation of intraocular pressure. The anterior chamber flare, the corneal endothelial cell density, the percentage of hexagonal cells and the corneal thickness showed no statistically significant difference between these two groups before and after surgery. CONCLUSION: Indocyaine green dye staining is a safe and useful technique in the white cataract phacoemulsification. We suggested to use ICG staining technique in the white cataract phacoemulsification.